Interleukin-2 fusion proteins and uses thereof

Abstract

The present invention generally relates to fusion proteins of immunoglobulins and interleukin-2 (IL-2). In addition, the present invention relates to polynucleotides encoding such fusion proteins, and vectors and host cells comprising such polynucleotides. The invention further relates to methods for producing the fusion proteins of the invention, and to methods of using them in the treatment of disease.

1 . A fusion protein comprising (i) an immunoglobulin molecule comprising a modification reducing binding affinity of the immunoglobulin molecule to an Fc receptor as compared to a corresponding immunoglobulin molecule without said modification, and (ii) two interleukin-2 (IL-2) molecules. 
     
     
         2 . The fusion protein of  claim 1 , wherein said immunoglobulin molecule is an IgG-class immunoglobulin molecule. 
     
     
         3 . The fusion protein of  claim 2 , wherein said IgG-class immunoglobulin molecule is IgG 1 . 
     
     
         4 . The fusion protein of  claim 1 , wherein said immunoglobulin molecule is a human immunoglobulin molecule. 
     
     
         5 . The fusion protein of  claim 1 , wherein said immunoglobulin molecule is not capable of specific binding to an antigen. 
     
     
         6 . The fusion protein of  claim 1 , wherein said immunoglobulin molecule comprises a heavy chain variable region sequence based on the human Vh3-23 germline sequence. 
     
     
         7 . The fusion protein of  claim 1 , wherein said immunoglobulin molecule comprises the heavy chain variable region sequence of SEQ ID NO: 9. 
     
     
         8 . The fusion protein of  claim 1 , wherein said immunoglobulin molecule comprises a light chain variable region sequence based on the human Vk3-20 germline sequence. 
     
     
         9 . The fusion protein of  claim 1 , wherein said immunoglobulin molecule comprises the light chain variable region sequence of SEQ ID NO: 11. 
     
     
         10 . The fusion protein of  claim 1 , wherein said Fc receptor is an Fcγ receptor. 
     
     
         11 . The fusion protein of  claim 1 , wherein said Fcγ receptor is a human Fcγ receptor. 
     
     
         12 . The fusion protein of  claim 11 , wherein said Fcγ receptor is a human Fcγ receptor. 
     
     
         13 . The fusion protein of  claim 1 , wherein said Fc receptor is an activating Fc receptor. 
     
     
         14 . The fusion protein of  claim 13 , wherein said Fc receptor is an activating Fc receptor. 
     
     
         15 . The fusion protein of  claim 1 , wherein said Fc receptor is selected from the group of FcγRIIIa (CD16a), FcγRI (CD64), FcγRIIa (CD32) and FcαRI (CD89). 
     
     
         16 . The fusion protein of  claim 1 , wherein said Fc receptor is human FcγRIIIa. 
     
     
         17 . The fusion protein of  claim 16 , wherein said Fc receptors is human FcγRIIIa. 
     
     
         18 . The fusion protein of  claim 1 , wherein said immunoglobulin molecule comprises an amino acid substitution at position 329 of the immunoglobulin heavy chains. 
     
     
         19 . The fusion protein of  claim 18 , wherein said amino acid substitution is P329G. 
     
     
         20 . The fusion protein of  claim 1 , wherein said immunoglobulin molecule comprises amino acid substitutions at positions 234 and 235 of the immunoglobulin heavy chains. 
     
     
         21 . The fusion protein of  claim 20 , wherein said amino acid substitutions are L234A and L235A (LALA). 
     
     
         22 . The fusion protein of  claim 1 , wherein said immunoglobulin molecule comprises the amino acid substitutions L234A, L235A and P3290 in the immunoglobulin heavy chains. 
     
     
         23 . The fusion protein of  claim 1 , wherein said IL-2 molecules are wild-type IL-2 molecules. 
     
     
         24 . The fusion protein of  claim 1 , wherein said IL-2 molecules are human IL-2 molecules. 
     
     
         25 . The fusion protein of  claim 1 , wherein said IL-2 molecules comprise the sequence of SEQ ID NO: 1 or SEQ ID NO: 3, particularly the sequence of SEQ ID NO: 3. 
     
     
         26 . The fusion protein of  claim 1 , wherein said IL-2 molecules are each fused at their N-terminal amino acid to the C-terminal amino acid of one of the immunoglobulin heavy chains of said immunoglobulin molecule, optionally through a peptide linker. 
     
     
         27 . The fusion protein of  claim 1 , wherein the fusion protein comprises the polypeptide sequences of SEQ ID NO: 17 and SEQ ID NO: 19. 
     
     
         28 . A polynucleotide encoding the fusion protein of  claim 1 . 
     
     
         29 . An expression vector, comprising the polynucleotide of  claim 28 . 
     
     
         30 . A host cell comprising the expression vector of  claim 29 . 
     
     
         31 . A method for producing a fusion protein, said method comprising the steps of: (i) culturing a host cell containing a polynucleotide encoding the fusion protein of  claim 1 , under conditions suitable for expression of the fusion protein; and (ii) recovering the fusion protein. 
     
     
         32 . A fusion protein comprising: (i) an immunoglobulin molecule comprising: a modification reducing binding affinity of the immunoglobulin molecule to an Fc receptor as compared to a corresponding immunoglobulin molecule without said modification, and (ii) two interleukin-2 (IL-2) molecules produced by the method of  claim 31 . 
     
     
         33 . A pharmaceutical composition comprising the fusion protein of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         34 . A pharmaceutical composition comprising the fusion protein of  claim 32  and a pharmaceutically acceptable carrier. 
     
     
         35 . A method for treating or preventing an autoimmune disease, said method comprising: administering the pharmaceutical composition of  claim 33  to a patient. 
     
     
         36 . The method of  claim 35 , wherein said autoimmune disease is selected from the group of type 1 diabetes, systemic lupus erythematosus, Crohn's disease and multiple sclerosis. 
     
     
         37 . A method for treating or preventing transplant rejection or graft-versus-host disease said method comprising: administering the pharmaceutical composition of  claim 33  to a patient. 
     
     
         38 . A method of treating a disease in an individual, said method comprising: administering to said individual a therapeutically effective amount of a composition comprising the fusion protein of  claim 1  in a pharmaceutically acceptable form. 
     
     
         39 . The method of  claim 38 , wherein said disease is an autoimmune disease. 
     
     
         40 . The method of  claim 39 , wherein said autoimmune disease is selected from the group of type 1 diabetes, systemic lupus erythematosus, Crohn's disease and multiple sclerosis. 
     
     
         41 . The method of  claim 39 , wherein said disease is transplant rejection or graft-versus-host disease. 
     
     
         42 . A method for selectively activating regulatory T cells in vitro or in vivo, said method comprising: administering a fusion protein of  claim 1 . 
     
     
         43 . The method of  claim 42 , wherein said activation comprises induction of proliferation and/or induction of IL-2 receptor signaling. 
     
     
         44 . The method of  claim 42 , wherein said method is in vitro and said fusion protein is used at a concentration of about 1 ng/mL or less, particularly about 0.1 ng/mL or less. 
     
     
         45 . The method of  claim 42 , wherein said method is in vivo and said fusion protein is used at a dose of about 20 μg/kg body weight or less. 
     
     
         46 . The method of  claim 45 , wherein said dose is less than or equal to about 12 μg/kg body weight.